brucellosis is a zoonotic disease transmitted to humans either from animals or from their products.although brucellosis can be found worldwide, the mediterranean basin, south and central america, easterneurope, asia, africa, the caribbean, and the middle east have currently been listed as high-risk regions. thegenus brucella is classified in at least nine species. brucella melitensis is the glo...

Hyperthermophile Pyrococcus furiosus grows optimally near 100°C and is an important resource of many industrial and molecular biological enzymes. To study the structure and function of P. furiosus proteins at whole genome level, we constructed expression plasmids of each P. furiosus gene using a ligase-independent cloning method, which was based on amplifying target gene and vector by PCR using...

The ability to assemble DNA sequences de novo through efficient and powerful DNA fabrication methods is one of the foundational technologies of synthetic biology. Gene synthesis, in particular, has been considered the main driver for the emergence of this new scientific discipline. Here we describe RapGene, a rapid gene assembly technique which was successfully tested for the synthesis and clon...

The nested polymerase chain reaction (PCR) method was used for the amplification of the influenza A H1N1 virus hemagglutinin monoclonal antibody light-chain and heavy-chain genes. Sequence analysis of the obtained genes was then used to identify common cloning methods of the mouse immunoglobulin-kappa (Igκ) light-chain and heavy-chain variable gene regions. Twenty-two pairs of amplification pri...

Background: In reverse genetics studies, minigenomes of negative sense RNA viruses are used to model the process virus replication and evaluate rescue system in course construction recombinant virus. current study, a minigenome (MG) for lentogenic D58 vaccine strain Avian orthoavulavirus 1 (AOAV-1) (Newcastle disease virus) was developed establish genetic this strain. Methods: The constructed b...

With the increasing number of polymorphic markers identified on the genetic map, positional cloning has become an extremely powerful tool, permitting the linkage and identification of a large number of disease genes. As the human genome project pro­ gresses and the gene map becomes as dense as the microsatellite polymorphism map, scientists will identify more disease genes through a combination...

The replication region of the Bacillus thuringiensis plasmid, pHT1030, was treated with hydroxylamine. Various copy-number mutants were selected and subsequently used to construct shuttle vectors with multiple cloning sites. These recombinant plasmids are very stable and allowed the cloning of a delta-endotoxin-encoding gene in B. thuringiensis. Comparison between gene expression level and vect...

background: acanthamoeba spp. is the causative agent of blindness keratitis and fatal encephalaitis. presence of acanthamoeba spp. in a wide variety of niches such as different water types can lead to exposure of high risk people such as contact lens wearers. the main aim of the present study was to explore the occurrence of acanthamoeba genotypes in the recreational water sources using both mo...

We have used the ability to induce high levels of Ty transposition to develop a method for transposon mutagenesis in Saccharomyces cerevisiae. To facilitate genetic and molecular analysis, we have constructed GAL1-promoted TyH3 or Ty917 elements that contain unique cloning sites, and marked these elements with selectable genes. These genes include the yeast HIS3 gene, and the plasmid PiAN7 cont...

background: the hbv-x (hbx) protein is believed to contribute to the development of hcc. however, the molecular mechanisms involved in hbx-mediated hepatocarcinogenesis remain obscure. in this study, the effect of hepatitis b virus x gene and its protein product hbxag on expression of p53 gene in hep g2 cell line was investigated. methods: viral dna extracted from hbv-positive serum and hbx gen...